1. Charakterystyka żywności / składnika

Introduction on the process used for characterisation of food constituents that are microorganisms: Microorganisms or microbes (e.g. bacteria) are living organisms, and can change over time depending on culture conditions. Correct identification of the microorganism‟s species and strain is of critical importance, as the observed effects are species and strain specific. The appropriate classification, identification and nomenclature of microorganisms constitute the starting point for the assessment of microbial properties. Classification assigns an organism to a known taxonomic group according to its similarity to that group. This allows the prediction of the properties of the microorganism on the basis of what is already known about the taxa. A reliable identification confirms the identity of the strain(s) used in a given process and requires the use of appropriate methods. Traditional phenotypic identification of bacteria is not always reliable since certain species cannot be distinguished by these methods. Molecular techniques have emerged in recent years as a replacement or complement to traditional phenotypic tests. DNA-DNA hybridization has become the generally accepted standard for determination of bacterial species identification. However this technique is difficult to perform and requires an expertise not normally present in the food industry. For these reasons phylogenetically based approaches such as sequence analysis of the 16S rRNA gene has proven to be a useful tool for bacterial identification. The EU-funded PROSAFE project concluded that biochemical tests should not be used as a stand-alone approach for identification of bacterial cultures (Vankerckhoven et al., 2008). The use of 16S rRNA gene sequence analysis was considered the best tool for routine species identification. Moreover, the use of sequence-based methods, such as 16S rRNA gene sequencing, was encouraged given their high reproducibility and data exchangeability (Vankerckhoven et al., 2008). The FAO/WHO expert group (FAO, 2006) recommends that phenotypic tests should be done first, followed by genetic identification, using methods such as DNA-DNA hybridization or 16S rRNA sequence analysis. Nevertheless, it is important to underline that in some cases 16S rRNA sequencing has a limited resolution and it may not be enough for discrimination of closely related species (Felis and Dellaglio, 2007; Vankerckhoven et al., 2008) being necessary to use other methods.
For the strain identification (characterisation of the strain by genetic typing), the FAO/WHO working group also recommended that strain typing has to be performed with a reproducible genetic method or using a unique phenotypic trait (FAO, 2006). DNA macrorestriction followed by Pulsed Field Gel Electrophoresis (PFGE) is considered as the generally accepted standard (FAO, 2006) and it has been extensively used for differentiating commercial microorganism strains. Other discriminatory molecular methods, such as Randomly Amplified Polymorphic DNA (RAPD) or Amplified rDNA restriction analysis (ARDRA) among others, are also available for strain characterisation. Hence, species identification and sufficient characterisation (genetic typing) at strain level by using internationally accepted molecular methods is needed. In addition, strains should be named according to the International Code of Nomenclature. Although there is no direct requirement on deposition of the particular strain in an internationally recognised culture collection, the FAO/WHO (FAO, 2006) recommends that strains should also be deposited in an internationally recognised culture collection (with access number). These will assure the tracking and access of scientists and regulatory authorities to the strain and related information in case it is needed. In the context of the Regulation (EC) nº 1924/2006, the purposes of characterisation are to confirm the identity of the food/constituent that is the subject of the health claim, and to establish that the studies provided for substantiation of the health claim were performed with the food/constituent in respect of which the health claim is made. Although not required for substantiation of a claim, characterisation should also be sufficient to allow control authorities to verify that the food/constituent which bears a health claim is the same one that was the subject of a community authorisation. The Panel has decided to use the following criteria for characterisation of food constituents that are microorganisms, which are the subject of health claims:
 Species identification by DNA-DNA hybridization or 16S rRNA sequence analysis.
 Strain identification by DNA macrorestriction followed by PFGE, RAPD, ARDRA or other internationally accepted genetic typing molecular methods.
Only when these two criteria were fulfilled, the microorganism was considered to be sufficiently characterised. In case of combination of several microorganisms, the Panel considers that if one microorganism used in the combination is not sufficiently characterised, the combination proposed is not sufficiently characterised. The characterisation of food constituents that are microorganisms, which are the subject of health claims pursuant to Article 13 of the Regulation (EC) nº 1924/2006, is based on evaluation of available references up to 31 December 2008, including the following:
 The information provided by the Member States in the consolidated list of Article 13 health claims and references that EFSA has received from Member States or directly from stakeholders;
 Generally available data obtained by searching PubMed and Web of Science databases by using the strain name as search term.

1.4. Characterisation of a combination of “Lactobacillus acidophilus ATCC SD5221 and Bifidobacterium lactis ATCC SD5220” (ID 869)

The food constituent that is the subject of the health claim is a combination of “Lactobacillus acidophilus ATCC SD5221 and Bifidobacterium lactis ATCC SD5220”. Lactobacillus acidophilus ATCC SD5221 (hereafter L. acidophilus ATCC SD5221) - The strain L. acidophilus ATCC SD5221, also known as L. acidophilus NCFM, species identity as well as the strain identity and characteristics have been clearly established by using both phenotypic and genotypic methods (Sanders and Klaenhammer, 2001). In addition, the genome L. acidophilus NCFM (ATCC SD5221) is publicly available at the National Center for Biotechnology Information (NCBI) database (Altermann et al., 2005) (accession nº NC_006814). The Panel considers that L. acidophilus ATCC SD5221, which is the subject of the health claim ID 869, is sufficiently characterised. A culture collection number from the American Type Culture Collection (ATCC) is provided. Bifidobacterium lactis ATCC SD5220 (hereafter B. lactis ATCC SD5220) - The identification/characterisation of the strain B. lactis ATCC SD5220, also known as B. lactis Bi-07, is not included in the studies provided as reference material. Some phenotypic features of the strain are known (Ding and Shah, 2007), but no information regarding genotypic identification/characterisation was found in the literature. The species B. lactis has been reclassified as B. animalis ssp. Lactis (Masco et al. 2004). It is important to point out that it may not be possible to differentiate the commercially available B. animalis ssp. lactis strains from each other on the basis of traditional genetic methods (e.g. PFGE) (Engel et al. 2003; Gueimonde et al., 2004) and may be necessary to use multi-locus sequencing or genome-wide approaches. The Panel considers that B. lactis ATCC SD5220, which is the subject of the health claim ID 869, is not sufficiently characterised. A culture collection number from the American Type Culture Collection (ATCC) is provided. The Panel considers that the combination of “Lactobacillus acidophilus ATCC SD5221 and Bifidobacterium lactis ATCC SD5220”, which is the subject of the health claim ID 869, is not sufficiently characterised.

Oświadczenia zdrowotne

ID 869 - Lactobacillus acidophilus (ATCC SD5221), Bifidobacterium lactis ATCC SD5220

1. Charakterystyka żywności / składnika

1.4. Characterisation of a combination of “Lactobacillus acidophilus ATCC SD5221 and Bifidobacterium lactis ATCC SD5220” (ID 869)

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